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目的:探讨高剂量X射线照射前后低氧诱导因子-1α(HIF-1α)在人鼻咽鳞癌CNE1细胞株中的表达情况。方法:体外培养人鼻咽鳞癌CNE1细胞株,待细胞进入对数生长期后行6MV X射线照射,照射方法为2Gy/次,隔日1次,共25次,累积剂量50Gy,收集照射前后细胞采用免疫荧光染色、RT-PCR和蛋白质印迹法检测高剂量X线照射前后CNE1细胞株中HIF-1α基因及其蛋白的表达。结果:照射前后HIF-1α基因的表达的半定量值分别为0.23±0.02和0.37±0.04,t=-5.422,P=0.006;其蛋白的表达的半定量值分别为0.05±0.01和0.08±0.01,t=-3.674,P=0.021;X线照射50Gy后HIF-1α在CNE1细胞中的表达比照前明显增高,P<0.05;共聚焦显微镜观察到X线照射50Gy后的CNE1细胞中绿色荧光强度高于照射前。结论:X射线照射可以引起HIF-1α在CNE1细胞中表达升高。
Objective: To investigate the expression of hypoxia inducible factor-1α (HIF-1α) in human nasopharyngeal squamous cell carcinoma CNE1 cell line before and after high-dose X-ray irradiation. Methods: Human nasopharyngeal squamous cell carcinoma cell line CNE1 was cultured in vitro. When the cells entered the logarithmic growth phase, they were irradiated with 6MV X-ray. The irradiation method was 2Gy / time, every other day for 25 times, the cumulative dose was 50Gy. Immunofluorescence staining, RT-PCR and Western blotting were used to detect the expression of HIF-1α gene and its protein in CNE1 cell line before and after high-dose X-ray irradiation. Results: The semiquantitative values of HIF-1α gene expression before and after irradiation were 0.23 ± 0.02 and 0.37 ± 0.04, respectively, t = -5.422 and P = 0.006. The semiquantitative values of HIF-1α gene expression were 0.05 ± 0.01 and 0.08 ± 0.01 , t = -3.674, P = 0.021. The expression of HIF-1α in CNE1 cells after X-ray irradiation was significantly higher than that before the irradiation, P <0.05. The green fluorescence intensity of CNE1 cells after 50Gy X-ray irradiation was observed by confocal microscopy Higher than before irradiation. Conclusion: X-ray irradiation can induce the expression of HIF-1α in CNE1 cells.